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Cat. No.: T4BGT-500 (for 500U)

Cat. No.: T4BGT-2500 (for 2500U)

Description

T4 Phage β-Glucosyltransferase (T4-BGT), also known as T4 β-glucosyltransferase, catalyzes the transfer of glucose moieties from uridine diphosphate glucose (UDP-glucose) to 5-hydroxymethylcytosine (5-hmC) residues in double-stranded DNA, forming β-glucosyl-5-hydroxymethylcytosine (5-gmC). DNA containing 5-gmC is resistant to cleavage by restriction endonucleases such as MunI and MfeI, thereby protecting 5-hmC from enzymatic digestion.

T4-BGT is widely used in next-generation sequencing (NGS)-based methylation profiling methods, including:

• Enzymatic methyl sequencing (EM-seq)

• APOBEC-coupled epigenetic sequencing (ACE-seq)

• TET-assisted bisulfite sequencing (TAB-seq)

Whole genome bisulfite sequencing (WGBS) has long been considered the gold standard for mapping DNA methylation. However, bisulfite treatment can damage DNA, leading to fragmentation, loss of material, and GC bias in methylated regions. Enzyme-based methylation sequencing methods overcome these limitations.

In EM-seq, whole-genome methylation library preparation involves two enzymatic steps:

1. TET2 (Tet methylcytosine dioxygenase 2) and T4-BGT convert 5-methylcytosine (5-mC) and 5-hydroxymethylcytosine (5-hmC) into 5-carboxylcytosine (5-caC) and β-glucosyl-5-hydroxymethylcytosine (5-gmC), respectively. These modified cytosines are resistant to deamination by APOBEC3A.

2. APOBEC3A, a cytidine deaminase, selectively deaminates unmodified cytosines (C) to uracil (U), without affecting 5-caC or 5-gmC. This enables clear differentiation between modified and unmodified cytosines.

By combining TET2, T4-BGT, and APOBEC3A, EM-seq achieves specific detection of 5-mC and 5-hmC, offering a robust alternative to traditional bisulfite sequencing.

Applications

• Specific detection of 5-hydroxymethylcytosine (5-hmC) in DNA using methods such as EM-seq, ACE-seq, and TAB-seq
• Enrichment of DNA fragments containing 5-hmC
• Radiolabeling of 5-hmC residues using radioactive UDP-glucose as a donor substrate

 Source

Recombinant T4-BGT gene expressed in Escherichia coli and purified to high quality.

Purity

• Free of DNase (endonuclease and exonuclease activity)
• Free of RNase
• Free of phosphatase
• Purity >95%  

Inactivation

T4-BGT is inactivated by heating at 65°C for 10 minutes.

Storage 

Store at –20ºC. Stable for at least two years under recommended conditions.

Precautions 

• Keep the product on ice or in an ice bath during use. Return to –20ºC storage immediately after use.
• This product is intended for use by qualified professionals in scientific research only. It is not for use in clinical diagnosis or treatment, food or pharmaceutical applications, and must not be stored in residential settings.
• For your safety and health, please wear a lab coat and disposable gloves when handling this product.

Related:

• Recombinant APOBEC3A (A3A)
• Recombinant Ten-Eleven Translocation 2 (TET2)
• T4 Phage β-Glucosyltransferase (T4-BGT)

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